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Biological sensor for detecting glutamic pyruvic transaminase

Alanine aminotransferase and biosensor technology, applied in the direction of biological testing, microbial measurement/inspection, biochemical equipment and methods, etc., can solve the problems that affect the accuracy and stability of detection, are susceptible to external interference, and are difficult to detect with small currents to achieve high electron transfer efficiency, ensure consistency, and increase the effect of current response signals

Inactive Publication Date: 2007-01-24
INST OF ELECTRONICS CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Because the concentration of alanine aminotransferase in the blood is relatively low, generally in tens of units, the biosensor based on the above technology has a small response current (pA-nA), a long response time (10 minutes, or even longer), and low sensitivity. lower disadvantage
It is difficult to detect small currents, especially with portable instruments, which are easily disturbed by the outside world, thus affecting the accuracy and stability of detection
Due to the relatively long response time, blood coagulation will also bring certain difficulties to the test

Method used

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  • Biological sensor for detecting glutamic pyruvic transaminase
  • Biological sensor for detecting glutamic pyruvic transaminase
  • Biological sensor for detecting glutamic pyruvic transaminase

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0033] Embodiment one: if figure 1 and figure 2 As shown, the biosensor for detecting alanine aminotransferase of the present invention comprises an insulating substrate 10, which is a polycarbonate sheet with a thickness of 0.5 mm, on which a layer of protective plastic film is pasted by electrostatic force, and a A high-precision film engraving machine engraves the pattern of the masking mask of the thin-film electrode required, and deposits a gold film on the substrate 10 through the metal film deposition process, with a thickness of about 100nm. After removing the masking mask, a flat metal film can be formed Electrode, one end of the film electrode is used as the access end of the insertion detection instrument: access end 7, access end 8 and access end 9, and a spacer 4 with a thickness of 0.125mm is pasted on the electrode surface of the other end, spacer 4 A reaction channel 5 (4.0mm×8.0mm) is engraved on it, and the electrode systems in the reaction channel 5 are re...

Embodiment 2

[0037] Example 2: 18 μl of alanine aminotransferase (GPT) reaction reagent was added to a 0.5 ml centrifuge tube. The reagent contains 0.01M disodium hydrogen phosphate-sodium dihydrogen phosphate with a pH value of 7.4. The buffer contains 0.1% triton (TX-100), 2U / ml glutamic acid oxidase, 10mM alginic acid Sodium, 10mg carboxymethyl cellulose (CMC) is dissolved and made the mixed solution. Then add 2 μl of alanine aminotransferase (GPT) solution of different concentrations. After 2 minutes of reaction, add the solution in the centrifuge tube dropwise to the electrode in the reaction channel 5 of the sensor, apply 0.0V voltage across the working electrode 2 and reference electrode 1, and read the reaction value after 60 seconds. current. Each GPT concentration was measured three times with different sensors.

[0038] The sensor adopts the above-mentioned manufacturing method, but no reaction reagent is added. Experimental results such as image 3 , when the concentration...

Embodiment 3

[0039] Example 3: 20 μl of alanine aminotransferase (GPT) reaction reagent was added to a 0.5 ml centrifuge tube. The reagent contains 0.01M disodium hydrogen phosphate-sodium dihydrogen phosphate with a pH value of 7.4. The buffer contains 0.1% triton (TX-100), 2U / ml glutamic acid oxidase, 10mM alginic acid Sodium, 10mg carboxymethyl cellulose (CMC) is dissolved and made the mixed solution. Dry in a constant temperature drying oven at 35°C for 24 hours. Take 20 μl of human serum sample and add it to a centrifuge tube containing dry reagents, stir well, and after 2 minutes of reaction, add the solution in the centrifuge tube dropwise to the electrode in the reaction channel 5 of the sensor, between the working electrode 2 and the reference electrode 1. Apply a voltage of 0.0V to both ends, and read the corresponding current after 60 seconds. response curve as Figure 4 . The response current of 3U / l GPT is 39.8nA, and the response current of 112U / l GPT is 104.9nA, thus, th...

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Abstract

The present invention is a kind of biosensor for quickly detecting glutamate-pyruvate transaminase. The biosensor is made through micro electromechanical processing process to form metal film electrodes and modifying the surface of the film electrodes with one layer of oxidation reduction polymer film as electronic medium to transfer electron between enzyme and electrode surface. The biosensor of the present invention has short response time and high sensitivity, and may be used in the quick detection of glutamate-pyruvate transaminase in fluid medium, such as blood, blood serum, urea, etc.

Description

technical field [0001] The invention relates to a biosensor for rapid detection of alanine aminotransferase, a manufacturing method and detection technology for mass production thereof. Background technique [0002] Alanine aminotransferase (GPT) is one of the most active transaminases in the body. It is widely distributed in various tissues and organs of the human body, especially in the liver, which is 3 times that of the heart, muscle and kidney, and nearly 3,000 times that of serum. . When organs such as the liver are damaged, intracellular GPT is released into the blood, which increases the activity of GPT in the blood. Therefore, the increase of GPT in the blood is almost always caused by liver disease, and the GPT in the blood is usually used as an important indicator to judge whether the liver function is normal. At present, GPT in blood is mainly performed on large-scale blood biochemical analyzers. Due to the need to use special devices, the measurement requires ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/52G01N33/50
Inventor 李华清蔡新霞崔大付刘春秀姜利英郭宗慧
Owner INST OF ELECTRONICS CHINESE ACAD OF SCI
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